Immunology and Virology

PRACTICAL ABO-Rh Blood Typing

Objectives
1. Perform standard test used for blood type identification
2. Understand the importance of blood type identification and its uses
3. Understand the concept of haemagglutination

Materials
1. Blood Test Card
2. Blood
3. Anti-Rh (D) serum
4. Anti-A serum
5. Anti-B serum
6. Mixing sticks (blue, yellow, white)

Methodology
1. A Blood Test Card (or a glass slide) is obtained. A drop of Anti-A serum (blue) is placed in
the circle labeled “Anti-A”. The cap is replaced on the Anti-A vial. The cap is always
replaced on one vial before opening the next vial to prevent cross contamination.

2. A drop of Anti-B serum (yellow) is placed I the circle labeled “ Anti-B” and then the cap is
replaced.

3. If the Rh typing is being done (if not, proceed to step 4), a drop of Anti-D serum (clear) is
placed in the circle labeled “Anti-D”. The cap is replaced on the Anti-D vial.

4. The tip of middle finger is scrubbed with an alcohol-soaked swab (or if blood cells are
supplied, a drop of blood cells are put in each “Blood” circle and proceeded to step 8.

5. A disposable sterile lancet, is removed from its wrapper. The middle finger is firmly
pressed about 1 cm from the fingertip with the thumb of the same hand and the clean
fingertip is punctured with the lancet. The lancet is discarded. A lancet should never be
reused; someone else’s lancet should never be reused,

6. The finger is gently pressed to force out a drop of blood onto each of the circles labeled
“Blood” on the Blood Test Card.

7. Any excess blood from the finger is wiped with the alcohol swab. In most cases, bleeding
will stop 1 to 2 minutes. Should bleeding continue, the finger is covered with an
adhesive bandage.

8. If Rh typing are being done (if not, proceeded to step 8), the white stirring stick is used
to mix one drop of blood with the Anti-D serum. The blood and the serum mixture are
carefully spread with the stirring stick until the mixture covers both connecting circles.
The stirring stick is then discarded.
 Immunology and Virology

9. The blue stirring stick is used to mix one drop of blood with the Anti-A serum. The blood
and the serum mixture is carefully spread with the stirring stick until both connecting
circles are covered by the mixture. Then the stirring stick is discarded.

10. The yellow stirring stick is used to mix one drop of blood with the Anti-B serum. The
blood and the serum mixture is carefully spread with the stirring stick until both
connecting circles are covered by the mixture. Then the stirring stick is discarded.
Note: Careful precautions are taken to ensure no cross contamination occurs as even
the slightest mixing will ruin the test results.

11. The test card is gently rocked back and forth for 1 minute. Each mixture is ensured to be
in its circle.

12. The Blood Test Card is tilted, allowing he blood and the serum mixtures to drain to the
side but not out of the circles. The appearance of thin blood films are compared with
the illustration table in the manual. If a film remains uniform in appearance, there is no
agglutination. If a film appears granular, agglutination has occurred. (Positive Rh
reactions may take longer time than positive A and B reactions. Five minutes may be
required to obtain an accurate Rh reading).

13. If the test results are not clear, steps 11 and 12 are repeated.

14. After the test sample is dry, a piece of cellophane tape is used to cover it.

15. The blood type is recorded on the Blood Test Card.

Result

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Discussion

1. Describe the components of blood.

Component of Blood Description Function

Red blood cells; biconcave
Erythrocytes
discs without nuclei
White blood cells;
Leukocytes
nucleated phagocytic cells
Platelets; small
Thrombocytes
cytoplasmic fragments
Plasma Fluid portion of blood
Transport O2 and CO2
Provide immune responses
Responsible for blood
clotting
Transporting nutrients,
gases, and vitamins;
regulating electrolyte and
fluid balance; maintain a
consistent blood pH

2. Explain how does red blood cell (RBC) can form agglutination when mix with the specific
antibody against the correspond RBC?
Agglutination occurs due to the reaction that is caused through the interaction of the
proteins in the red blood cells and the antibodies present in the blood plasma. Example:
Antibodies-b, also known as Anti-a sera, will clump red blood cells containing A-antigens, or
type A blood. Anti-b sera will clump type B blood. Clumping will occur in both sera with type
AB blood and in neither sera with type O blood.

3. Describe about Rhesus factor and complication to the fetus when the mother carry the Rh+
antigen.
Rh factor is an antigen, a substance that stimulates the production of antibodies to fight
foreign invaders, such as viruses, bacteria and transplanted organs. A given individual either
 Immunology and Virology

has the antigen already in their blood (they are Rh positive), or they do not (they are Rh
negative). A patient's Rh status effects how he or she handles blood transfusions or organ
transplants.

Rh-positive women does not having risk to have children with the Rhesus factor disease.

As for woman with Rh-; they will not encounter any problem with their first child. The
problem may arise after the birth of her first child. When the umbilical cord has been cut,
and if the baby has the Rh+, little of the baby's blood will flow back into the mother's body.
The mother's body will then become immune towards Rh+. When this happens, the
subsequent pregnancy for Rh+ baby will not succeed as the baby may die prematurely in the
womb or the mother undergoes miscarriages. But, if a Rh- woman is carrying a Rh- baby, it
will not be a problem for the mother and the baby in her womb and also for the woman's
subsequent pregnancy.

Conclusion
Through this experiment, we become familiar with standard test used for blood type
identification; understand and appreciate the concept and importance of blood type
identification and its uses.